Correlation Analysis Between Parkinson’s Disease And Intestinal Flora

BackgroundParkinson’s disease(PD),as the second most common neurodegenerative disease in the world,seriously affects the health and quality of life of patients,and the etiology has not yet been fully clarified.Mounting number of studies have found that gut microbes exert an important part in its occurrence and development.It plays a crucial role,and the changes in the structure and the relative abundances of intestinal flora affect the process of PD in quite a few aspects such as "neurology-immunity-inflammation",so it is a potential diagnostic biomarker and innovative target for intervention.The study of the correlation between flora and Parkinson’s disease patients is of great significance for the diagnosis and treatment of PD.ObjectiveThe main objective of this research is to explore the gut biomarkers of Parkinson’s disease,the gut flora identification markers of different motor subtypes,and new targets for relevant treatments,to provide new evidence for the diagnosis,prevention,and future treatment of PD and feasible strategies.M ethodsIn this study,38 patients with idiopathic Parkinson’s disease(Parkinson’s disease group)and 23 age and sex matched control subjects(non-Parkinson’s disease group included 15 healthy controls,8 recognized cognitive impairment patients),first collect general clinical data,serologic indicators,motor symptoms and non-motor symptoms scale evaluation of PD patients.We formulate 9 comparison plans based on clinical symptoms.At the same time,feces samples from all subjects were collected,bacterial genomic DNA was extracted,gene amplification and purification,gene library construction,and high-channel sequencing of 16 S r RNA V3+V4 variable regions were performed.For the annotation of intestinal flora,refer to Greengenes database.Using R software package,SPSS21.0 and other software,Kruskal-Wallis,ANOVA,Wilcoxon,ANCOM and other statistical methods were used to analyze the diversity and structural differences of the intestinal flora among the groups;the LEf Se method was used to analyze the intestinal flora markers were identified;Spearman correlation analysis and RDA method were used to analyze the correlation between the clinical manifestations of PD patients and the intestinal flora;finally,using PICRUSt2 software,based on the Meta Cyc database,the functional pathways with significant differences in the intestinal flora genome were predicted.Results1.Analysis of intestinal flora diversity showed that,Parkinson’s disease group and healthy control group,healthy control group and pure cognitive impairment group,motor subtype group pigd(abnormal posture gait disorder group),td(tremor-based group)and In the mix(mixed group),there was a significant statistical difference in the diversity index of intestinal flora among the groups(P<0.05),suggesting that there were significant differences in the structure of the intestinal flora among these groups.There was no significant difference in bacterial diversity between Parkinson’s disease group and pure cognitive impairment group(P>0.05).2.The analysis of the relative abundance difference of intestinal flora showed that compared with the healthy control group,in the Parkinson’s disease group,Actinobacteria,Peptococcaceae,Ruminococcus,Bifidobacterium,Oscillospira,Coprococcus,Akkermansia were significantly increased;The relative abundances of Gut Bacteria which decline significantly,include Bacteroidetes,Sphingomonadaceae,Sphenodons,Eubacterium,Bacteroides,Faecalibacterium,Prevotella,Roseburia,Blautia.They were all statistically significant(P<0.05).There were also significant differences in the relative abundance of gut microbiota between the 3 motor subtype subgroups(pigd,td,and mix)of Parkinson’s disease.Comparing the pure cognitive impairment group with the healthy control group,the changes in the bacterial flora of the pure cognitive impairment group were like those of the Parkinson’s disease group.3.LEf Se analysis shows that Parkinson’s disease patients were different from the healthy control group of the most important biomarkers,among which significantly increased were Oscillospira,Blautia,Coprococcus,corresponded to the hierarchy was Firmicutes,Ruminococcaceae,Oscillospira,guilliermondii;The Lower biomarkers of Parkinson’s disease patients are Firmicutes,Erysipelotrichaceae,Eubacterium,dolichum and Firmicutes,Lachnospiraceae,Roseburia,inulinivorans.Compared with the healthy control group,the markers that were significantly increased in the pure cognitive impairment group were Verrucomicrobia,Verrucomicrobiaceae,Akkermansia.The most important marker to distinguish between the three motor subtypes was,Bacteroidetes,Bacteroidaceae,Bacteroides,and annotated to species level,mix group is coprophilus,td group is uniformis,and pigd group is fragilis.The above differences were statistically significant(P<0.05).4.Correlation analysis showed the intestinal flora which showed significant changes in PD patients in the relative abundance difference analysis was also significantly correlated with the clinical manifestations and blood lipid indicators(TD,APOA1,HDL)of PD patients.Blautia was negatively correlated with HY grade,NMSS,and PDQ39;Eubacterium was positively correlated with HY grade;Roseburia was negatively correlated with GSRS score,bradykinesia-tonic score,and positively correlated with tremor score correlation;Ruminococcus was negatively correlated with NMSS,PDQ39 score,SDS standard score,HDL;Coprococcus was negatively correlated with most motor symptom scores(except tremor score)and PDQ39;Akkermansia was negatively correlated with MDS-UPDRS Ⅲ total score,bradykinesia-rigidity score,and limb symptom score were negatively correlated;Sphingomonas was negatively correlated with MMSE score,MOCA total score,and TLED.The above differences were statistically significant(P<0.05),and the absolute value of the correlation coefficient was >0.4.5.Gene function prediction showed that compared with healthy controls,the functions of the two Ltryptophan metabolic pathways(NADSYN-PWY and PWY-5651)in patients with Parkinson’s disease were significantly decreased;among the three motor subtypes,compared with the mix group,Taurine and hypotaurine metabolism function in pigd group and td group increased significantly,and it was more obvious in td;compared with td,the function of the Carotenoid biosynthesis function is increased significantly in pigd group and mix group,especially in the mix.The above differences were all statistically significant(P<0.001 and adjusted P<0.001).Conclusions1.The intestinal flora of patients with Parkinson’s disease is significantly different from that of healthy people,and we found six levels of characteristic biomarkers among the gut flora.2.There are significant differences in the intestinal flora among the three motor subtypes of Parkinson’s disease MIX type,PIGD type and TD type,and the subspecies of Bacteroides are the key markers to distinguish the tree types.3.The intestinal flora is significantly related to the clinical manifestations of Parkinson’s disease(motor symptoms,non-motor symptoms)and blood lipid indicators(TD,APOA1,HDL).